U.S. patent application number 10/172005 was filed with the patent office on 2003-09-11 for synergistic enhancement of cognitive ability.
This patent application is currently assigned to Neurologic, Inc.. Invention is credited to Alkon, Daniel L., Sun, Mlao-Kun.
Application Number | 20030171385 10/172005 |
Document ID | / |
Family ID | 27807419 |
Filed Date | 2003-09-11 |
United States Patent
Application |
20030171385 |
Kind Code |
A1 |
Alkon, Daniel L. ; et
al. |
September 11, 2003 |
Synergistic enhancement of cognitive ability
Abstract
The present invention relates to the combination of a
methylxanthine and a carbonic anhydrase activator to provide
synergistic effects. The invention further relates to the
improved/enhanced cognitive ability of individuals, particularly
those suffering from various disorders, such as Alzheimer's
Disease, stroke, hypoxia, general dementia, ADHD, mental
retardation, and "sun down" syndrome.
Inventors: |
Alkon, Daniel L.; (Bethesda,
MD) ; Sun, Mlao-Kun; (Gaithersburg, MD) |
Correspondence
Address: |
HUNTON & WILLIAMS
INTELLECTUAL PROPERTY DEPARTMENT
1900 K STREET, N.W.
SUITE 1200
WASHINGTON
DC
20006-1109
US
|
Assignee: |
Neurologic, Inc.
|
Family ID: |
27807419 |
Appl. No.: |
10/172005 |
Filed: |
June 17, 2002 |
Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
|
|
60362081 |
Mar 7, 2002 |
|
|
|
Current U.S.
Class: |
514/263.31 ;
514/263.32; 514/400; 514/567 |
Current CPC
Class: |
A61K 2300/00 20130101;
A61K 2300/00 20130101; A61K 2300/00 20130101; A61K 31/198 20130101;
A61K 31/4172 20130101; A61K 2300/00 20130101; A61K 2300/00
20130101; A61K 31/52 20130101; A61K 31/198 20130101; A61K 31/417
20130101; A61K 31/4172 20130101; A61K 31/135 20130101; A61K 31/135
20130101; A61P 25/28 20180101; A61K 31/52 20130101; A61K 31/417
20130101 |
Class at
Publication: |
514/263.31 ;
514/263.32; 514/567; 514/400 |
International
Class: |
A61K 031/522; A61K
031/4172; A61K 031/198 |
Claims
1. A method for enhancing cognitive ability comprising
administering to a subject a combination of a methylxanthine and a
carbonic anhydrase activator in an amount effective to enhance
cognitive ability of said subject in a pharmaceutically suitable
carrier.
2. The method of claim 1, wherein said carbonic anhydrase activator
is phenylalanine, imidazole, dopamine, noradrenaline, adrenaline,
histamine, histidine, or 5-HT.
3. The method of claim 2, wherein said carbonic anhydrase activator
is phenylalanine.
4. The method of claim 1, wherein said methylxanthine is
theophylline or caffeine.
5. The method of claim 1, wherein the cognitive ability enhanced is
learning, memory, or attention.
6. The method of claim 1, wherein the subject is a human
7. The method of claim 1, wherein the subject is non-human.
8. The method of claim 1, wherein said effective amount of said
combination is an amount to treat cognitive impairment of a
neurological condition or disorder.
9. The method of claim 8, wherein said neurological condition is
Alzheimer's disease, attention deficit hyperactivity disorder, "sun
down" syndrome, hypoxia, ischeamic stroke, mental retardation,
stroke, general dementia, multi-infarct dementia, the Lewy-body
variant of Alzheimer's disease with or without association with
Parkinson's disease, Creutzfeld-Jakob disease, or Korsakow's
disorder.
10. The method of claim 8, wherein said disorder is age-associated,
is consequent upon electro-convulsive therapy or is the result of
brain damage.
11. The method of claim 10, wherein said brain damage is caused by
stroke, an anesthetic accident, head trauma, hypoglycemia, carbon
monoxide poisoning, lithium intoxication or a vitamin
deficiency.
12. A method for treating a subject with Alzheimer's Disease
comprising administering to said subject a pharmaceutically
effective amount of a combination of a methylxanthine and a
carbonic anhydrase inhibitor.
13. A method for treating a subject with Alzheimer's Disease
comprising administering to said subject a pharmaceutically
effective amount of a combination of theophylline and
phenylalanine.
14. A method for enhancing cognitive ability of a subject
comprising administering to said subject a pharmaceutically
effective amount of a combination of a methylxanthine and a
carbonic anhydrase activator.
15. A method for enhancing cognitive ability of a subject
comprising administering to said subject a pharmaceutically
effective amount of a combination of theophylline and
phenylalanine.
16. A method for treating hypoxia comprising administering to a
subject a pharmaceutically effective amount of a combination of a
methylxanthine and a carbonic anhydrase activator.
17. A method for treating hypoxia comprising administering to a
subject a pharmaceutically effective amount of a combination of
theophylline and phenylalanine.
18. A method for providing a neuroprotective effect for cells which
suffer from a hypoxic event comprising administering to a subject
in need of such effect a pharmaceutically effective amount of a
combination of a methylxanthine and a carbonic anhydrase
activator.
19. A method for providing a neuroprotective effect for cells which
suffer from a hypoxic event comprising administering to a subject
in need of such effect a pharmaceutically effective amount of a
combination of theophylline and phenylalanine.
20. A composition comprising a combination of a methylxanthine and
a carbonic anhydrase activator in an amount effective to enhance
cognitive ability, and a pharmaceutically effective carrier.
21. The composition of claim 20, wherein said carbonic anhydrase
activator is phenylalanine, imidazole, dopamine, noradrenaline,
adrenaline, histamine, histidine, or 5-HT.
22. The composition of claim 20, wherein said methylxanthine is
theophylline or caffeine.
Description
PRIORITY OF INVENTION
[0001] This application claims priority under 35 U.S.C. .sctn.
119(e) from U.S. Provisional Application No. 60/362,081 filed Mar.
7, 2002.
BACKGROUND OF THE INVENTION
[0002] (i) Field of the Invention
[0003] The present invention relates to the cognitive enhancement
through the administration of synergistic drugs.
[0004] (ii) Background of the Invention
[0005] Various disorders and diseases exist which affect cognition.
Cognition can be generally described as including at least three
different components: attention, learning, and memory. Each of
these components and their respective levels affect the overall
level of a subject's cognitive ability. For instance, while
Alzheimer's Disease patients suffer from a loss of overall
cognition and thus deterioration of each of these characteristics,
it is the loss of memory that is most often associated with the
disease. In other diseases patients suffer from cognitive
impairment that is more predominately associated with different
characteristics of cognition, for instance Attention Deficit
Hyperactivity Disorder (ADHD), focuses on the individual's ability
to maintain an attentive state. Other conditions include general
dementias associated with other neurological diseases, aging, and
treatment of conditions that can cause deleterious effects on
mental capacity, such as cancer treatments, stroke/ischemia, and
mental retardation. The present invention is directed toward the
treatment of these and other similar disorders through the repair
or amelioration of the cognitive deficits or impairments.
[0006] Cognition disorders create a variety of problems for today's
society. Therefore, scientists have made efforts to develop
cognitive enhancers or cognition activators. The cognition
enhancers or activators that have been developed are generally
classified to include nootropics, vasodilators, metabolic
enhancers, psychostimulants, cholinergic agents, biogenic amines
drugs, and neuropeptides. Vasodilators and metabolic enhancers
(e.g. dihydroergotoxine) are mainly effective in the cognition
disorders induced by cerebral vessel ligation-ischemia; however,
they are ineffective in clinical use and with other types of
cognition disorders. Of the developed cognition enhancers,
typically only metabolic drugs are employed for clinical use, as
others are still in the investigation stage. Of the nootropics for
instance, piracetam activates the peripheral endocrine system,
which is not appropriate for Alzheimer's Disease due to the high
concentration of steroids produced in patients while tacrine, a
cholinergic agent, has a variety of side effects including
vomiting, diarrhea, and hepatotoxicity.
[0007] Ways to improve the cognitive abilities of diseased
individuals have been the subject of various studies. Recently the
cognitive state related to Alzheimer's Disease and different ways
to improve patient's memory have been the subject of various
approaches and strategies. In the case of Alzheimer's Disease,
efforts to improve cognition, typically through the cholinergic
pathways or though other brain transmitter pathways, have been
investigated. This approach relies on the inhibition of acetyl
cholinesterase enzymes through drug therapy. Acetyl cholinesterase
is a major brain enzyme and manipulating its levels can result in
various changes to other neurological functions and cause side
effects. Cholinesterase inhibitors only produce some symptomatic
improvement for a short time. Additionally, the use of cholinergic
inhibitors only produces an improvement in a fraction of the
Alzheimer's Disease patients with mid to moderate symptoms and is
thus only a useful treatment for a small portion of the overall
patient population. As a result, use of the cholinergic pathway for
treatment of cognitive impairment, particularly in Alzheimer's
Disease, has proven to be inadequate. Additionally, current
treatments for cognitive improvement are limited to specific
neurodegenerative diseases and have not proven effective in
treatment across a broad range of cognitive conditions.
[0008] With regard to normal and abnormal memory both K.sup.+ and
Ca.sup.2+ channels have been demonstrated to play key roles in
memory storage and recall. For instance, potassium channels have
been found to change during memory storage. (Etcheberrigaray, R.,
et al. (1992) Proceeding of the National Academy of Science
89:7184; Sanchez-Andres, J. V. and Alkon, D. L. (1991) Journal of
Neurobiology 65:796; Collin, C., et al. (1988) Biophysics Journal
55:955; Alkon, D. L., et al. (1985) Behavioral and Neural Biology
44:278; Alkon, D. L. (1984) Science 226:1037). This observation,
coupled with the almost universal symptom of memory loss in
Alzheimer's patients, led to the investigation of ion channel
function as a possible site of Alzheimer's Disease pathology,
modulation by PKC, and its overall effect on cognition.
[0009] There still exists a need for the development of methods for
the treatment for improved overall cognition, either through a
specific characteristic of cognitive ability or general cognition.
There also still exists a need for the development of methods for
the improvement of cognitive enhancement whether or not it is
related to a specific disease state or cognitive disorder. The
methods and compositions of the present invention are needed and
will greatly improve the clinical treatment for diminished
cognitive ability whether related to a specific neurodegenerative
disease, hypoxia, stroke or similar disorder. The methods and
compositions also provide treatment and/or enhancement of the
cognitive state.
SUMMARY OF THE INVENTION
[0010] The present invention relates to compounds, compositions,
and methods for the treatment of conditions associated with the
impairment of cognitive ability. In a preferred embodiment, the
present invention further relates to compounds, compositions and
methods for the treatment of conditions associated with
neurodegenerative diseases, such as Alzheimer's Disease, memory
dysfunction, and ischemia/stroke. Treatment provides for
improved/enhanced cognitive ability. In another embodiment the
present invention relates to compounds, compositions, and methods
for the improvement/enhancement of cognitive ability.
[0011] In another aspect the present invention relates to the
combination of a methylxanthine and carbonic anhydrase activators,
to alter or test distinct molecular cascades, either in vivo or in
vitro, in order to provide enhanced cognitive response. In a
preferred embodiment the carbonic anhydrase activator is
phenylalanine. In a preferred embodiment the methylxanthine is
selected from theophylline and caffeine. Enhanced cognitive
response, for example, can be employed in the treatment of
Alzheimer's Disease.
[0012] Another aspect of the present invention relates to a method
for treating conditions related to hypoxia and improving/enhancing
the cognitive state of the subject comprising administering to the
subject an effective amount of a composition combining a
methylxanthine and a carbonic anhydrase activator. In a preferred
embodiment the carbonic anhydrase activator is phenylalanine. In a
preferred embodiment the methylxanthine is selected from
theophylline and caffeine.
[0013] Another aspect of the present invention relates to a
composition for improving/enhancing cognitive ability comprising:
(i) an effective amount of a combination of a methylxanthine and a
carbonic anhydrase activator; and (ii) a pharmaceutically effective
carrier. In a preferred embodiment the carbonic anhydrase activator
is phenylalanine. In a preferred embodiment the methylxanthine is
selected from theophylline and caffeine. In a preferred embodiment
the composition is used to improve/enhance cognitive ability
associated with Alzheimer's Disease or stroke/ischemia. In another
embodiment, the combination is delivered to subjects or models of
Alzheimer's Disease or stroke/hypoxia.
[0014] In one embodiment of the invention the combination of a
methylxanthine and a carbonic anhydrase activator results in
improved cognitive abilities. In a preferred embodiment the
carbonic anhydrase activator is phenylalanine. In a preferred
embodiment the methylxanthine is selected from theophylline and
caffeine. In one embodiment the improved cognitive ability is
memory. In another embodiment the improved cognitive ability is
learning. In another embodiment the improved cognitive ability is
attention.
[0015] Another embodiment of the invention is a method of improving
cognitive ability through the combination of a methylxanthine and a
carbonic anhydrase activator. In another embodiment of the
invention the combination of a methylxanthine and a carbonic
anhydrase activator is delivered to "normal" subjects. In another
embodiment of the invention the combination of theophylline and a
carbonic anhydrase activator is delivered to subjects suffering
from a disease, deteriorating cognitive faculties, or
malfunctioning cognition. In a preferred embodiment the method is a
method for treating Alzheimer's Disease cognitive degeneration. In
a preferred embodiment the carbonic anhydrase activator is
phenylalanine. In a preferred embodiment the methylxanthine is
selected from theophylline and caffeine.
[0016] In a preferred embodiment of the invention the combination
of a methylxanthine and a carbonic anhydrase activator is
administered through oral and/or injectable forms including
intravenously and intraventricularly. In another embodiment the
combination may be administered through a sports drink or as a food
supplement. In a preferred embodiment the carbonic anhydrase
activator is phenylalanine. In a preferred embodiment the
methylxanthine is selected from theophylline and caffeine.
[0017] The present invention therefore provides methods of treating
impaired memory or a learning disorder in a subject, the method
comprising administering thereto a therapeutically effective amount
of a methylxanthine and a carbonic anhydrase activator. The
compounds can thus be used in the therapeutic treatment of clinical
conditions in which memory defects or impaired learning occur. In
this way memory and learning can be improved and the condition of
the subject can thereby be improved.
[0018] The present invention is also particularly suited to
administration, particularly oral administration, since the
combination of a methylxanthine (e.g. theophylline) and a carbonic
anhydrase activator would be associated with a specific blood brain
barrier transporter (BBB). In a preferred embodiment the
transporter is the BBB transporter for phenylalanine.
[0019] The compositions and methods have utility in treating
clinical conditions and disorders in which impaired memory or a
learning disorder occurs, either as a central feature or as an
associated symptom. Examples of such conditions which the present
compounds can be used to treat include Alzheimer's Disease,
multi-infarct dementia and the Lewy-body variant of Alzheimer's
Disease with or without association with Parkinson's Disease;
Creutzfeld-Jakob Disease, Korsakow's disorder, attention deficit
hyperactivity disorder, hypoxia, ischeamic stroke, mental
retardation, general dementia, and "sundown" syndrome.
[0020] The compositions and methods can also be used to treat
impaired memory or learning which is age-associated, is consequent
upon electro-convulsive therapy or which is the result of brain
damage caused, for example, by stroke, an anesthetic accident, head
trauma, hypoglycemia, carbon monoxide poisoning, lithium
intoxication or a vitamin deficiency.
[0021] The pharmaceutical compositions and methods according to the
invention are useful in the enhancement of cognition, prophylaxis
and/or treatment of cognition disorders, wherein cognition
disorders include, but are not limited to, disorders of learning
acquisition, memory consolidation, and retrieval, as described
herein.
BRIEF DESCRIPTION OF THE DRAWINGS
[0022] FIG. 1 illustrates the improved cognitive ability of treated
rats using the Morris Water Maze paradigm as compared to control
(phenylalanine) and theophylline alone treated rats.
Phenylalanine-theophylline (orally; 50 mg/kg phenylalanine +2 mg/kg
theophylline) or theophylline (2 mg/kg); 2 doses at 1 hr interval,
with the 2nd dose administered about 0.5 hr prior to the 1st
training trial of the day); 10 rats/group.
[0023] FIG. 2(a) and 2(b) illustrate the swimming time, in each
quadrant of the Morris Water Maze, for control rats and rats
treated with PhePheTheo, respectively.
[0024] FIG. 2(c) demonstrates the cumulative target quadrant ratio
for the Morris Water Maze.
DETAILED DESCRIPTION OF PREFERRED EMBODIMENTS
[0025] Memory loss and impaired learning ability are features of a
range of clinical conditions. For instance, loss of memory is the
most common symptom of dementia states including Alzheimer's
Disease. Memory defects also occur with other kinds of dementia
such as multi-infarct dementia (MID), a senile dementia caused by
cerebrovascular deficiency, and the Lewy-body variant of
Alzheimer's Disease with or without association with Parkinson's
disease, or Creutzfeld-Jakob disease. Loss of memory is a common
feature of brain-damaged patients. Brain damage may occur, for
example, after a classical stroke or as a result of an anaesthetic
accident, head trauma, hypoglycaemia, carbon monoxide poisoning,
lithium intoxication, vitamin (B1, thiamine and B12) deficiency, or
excessive alcohol use or Korsakow's disorder. Memory impairment may
furthermore be age-associated; the ability to recall information
such as names, places and words seems to decrease with increasing
age. Transient memory loss may also occur in patients, suffering
from a major depressive disorder, after electro-convulsive therapy
(ECT). Alzheimer's Disease is in fact the most important clinical
entity responsible for progressive dementia in ageing populations,
whereas hypoxia/stroke is responsible for significant memory
defects not related to neurological disorders.
[0026] Individuals with Alzheimer's Disease are characterized by
progressive memory impairments, loss of language and visuospatial
skills and behavior deficits (McKhann et al., 1986, Neurology,
34:939-944). The cognitive impairment of individuals with
Alzheimer's Disease is the result of degeneration of neuronal cells
located in the cerebral cortex, hippocampus, basal forebrain and
other brain regions. Histologic analyses of Alzheimer's Disease
brains obtained at autopsy demonstrated the presence of
neurofibrillary tangles (NFT) in perikarya and axons of
degenerating neurons, extracellular neuritic (senile) plaques, and
amyloid plaques inside and around some blood vessels of affected
brain regions. Neurofibrillary tangles are abnormal filamentous
structures containing fibers (about 10 nm in diameter) that are
paired in a helical fashion, therefore also called paired helical
filaments. Neuritic plaques are located at degenerating nerve
terminals (both axonal and dendritic), and contain a core compound
of amyloid protein fibers. In summary, certain neuropathological
features including intracellular neurofibrillary tangles, primarily
composed of cytoskeletal proteins, and extracellular parenchymal
and cerebrovascular amyloid, characterize Alzheimer's Disease.
Further, there are now methods in the art for distinguishing
between Alzheimer's patients, normal aged people, and people
suffering from other neurodegenerative diseases, such as
Parkinson's, Huntington's chorea, Wemicke-Korsakoff or
schizophrenia further described for instance in U.S. Pat. No.
5,580,748 and U.S. Pat. No. 6,080,582.
[0027] Hypoxia/ischemic stroke remains one of the most devastating
threats to humans and a challenge to neuropharmacologists. Because
of the extreme sensitivity of neural structures involved in memory,
especially the hippocampal CA1 pyramidal cells, to hypoxia and
ischemia, memory impairment is common after cerebral
hypoxia/ischemia, bypass surgery, or heart attack. Cognitive
decline is evident in more than half to as many as three-quarters
of patients at the time of discharge from hospitals after
coronary-artery bypass grafting, as well as, in patients with
chronic lung diseases or oropharyngeal abnormality.
Hypoxic/ischemic consequences consist mainly of three forms:
functional disruption, cellular injury, and delayed cell loss
through apoptosis or necrosis, depending on the severity of the
insult.
[0028] It is well established that functions of mammalian neurons
are sensitive to acute hypoxia. The brain is a metabolically very
active organ, but it contains virtually no O.sub.2 reserve. Upon a
sudden occlusion of brain circulation (ischemia), the brain is left
with an O.sub.2 content of about 0.2 ml/100 g and intracellular
energy stores, which can support and maintain cellular energy for 1
to 2 min and 37.degree. C. Cerbral hypoxia/ischemia, as occurs with
environmental limitations, insufficient blood flow (cerebrovascular
hemorrhage, brain tumor, vascular occlusion, or cardiac arrest, by
pass surgery), respiratory dysfunction (obstruction of the airway,
lung dysfunction, or neural control failure) or the use of some
toxic substances, results in a high incidence of memory deficits
and moderate-to-profound memory loss in humans. Irreversible damage
to brain tissue is cause by 10 minutes of severe hypoxia in vivo
and in vitro. However, episodes of transient hypoxia may be more
relevant to a gradual memory decline. This may be particularly true
following a brief hypoxic event or the continuous insult, which
occurs with neurodegnerative diseases, such as AD, or during normal
aging. Further, experiments demonstrate that induced hypoxic
synaptic arrest compromises the ability of brains to learn and
memorize.
[0029] Thus, a selective deficit in explicit memory functions is
associated with neuronal loss/damage. While memory consolidation
and processing is not limited to the hippocampus, the hippocampal
CA1 pyramidal cells are among the most sensitive to
hypoxic/ischemic damage. A major modulator of GABAergic inhibition
in the hippocampus, present in CA1 pyramidal cells, is carbonic
anhydrase. In humans and other species, including rats, the
hippocampus has a broad role in information processing associated
with memory, including spatial, declarative/relational, and
episodic types of memory. In mammals, the hippocampus, a major
component of the medical temporal lobe, mediates learning of
associations between environmental contexts and sensory stimuli.
Damage restricted to the hippocampus leads to deficits in cognitive
tasks, particularly in spatial learning and memory. The existence
of "place cells," pyramidal hippocampal neurons that fire when the
animal is in a particular location in its environment, or when it
receives a specific stimulus or performs a specific behavior in a
particular place, provides additional support for the crucial role
of the hippocampus in spatial cognition. Signal processing within
the hippocampal network, including transmission of a .theta. rhythm
from the septum to the hippocampus, is under strict control of
interneurons that release GABA. Memory abnormalities that
characterize the early stages of Alzheimer's Disease (AD) involve
multiple neurotransmitter deficits in the hippocampal formation. It
is known that alterations in synaptic spines and loss of dendrites
during aging are associated with a significant decline in carbonic
anhydrase in the brain and that this decline is even more dramatic
in brains of AD patients.
[0030] Hypoxia significantly reduced cholinergic theta activity in
rat CA1 field and intracellular theta in the CA1 pyramidal cells,
recorded in hippocampal slices. Research suggests that hypoxia
releases adenosine and produces an inhibition of synaptic
transmission and intracellular signal cascade(s) involved in
generation/maintenance of hippocampal CA1 theta activity. This is
supported by studies indicating that the hypoxic synaptic arrest is
prevented by blocking the adenosine A1 receptor and that spatial
learning and functional impairment of the hippocampal CA1 synaptic
plasticity are preventable by the adenosine A1 receptor antagonist,
DPCPX. Additionally, it has been demonstrated that blocking
adenosine A1 receptors prevent the impairment of spatial learning
and memory and synaptic plasticity in response to non-injury
hypoxic episodes.
[0031] Synapses are considered a critical site through which
memory-related events realize their functional expression, whether
the events involve changed gene expression and protein translation,
altered kinase activities, or modified signaling cascades.
Cognition and synaptic plasticity involve operational changes in
preexisting synapses, the growth of new synapses, and processes
that involve multiple synaptic transmitters and signaling
molecules. A few proteins have been implicated in associative
memory including Ca.sup.2+ calmodulin II kinases, protein kinase C,
calexcitin, a 22-kDa learning-associated Ca.sup.2 + binding
protein, and type II ryanodine receptors. Memories are thought to
be a result of lasting synaptic modification in the brain
structures related to information processing. Ca.sup.2+ signaling,
controlled by the endoplasmic reticulum (ER) and the plasma
membrane, is a critical factor that induces changes in synaptic
plasticity. Not only might neural activity control the amount of
Ca.sup.2+ stored in the ER, but Ca.sup.2+ can also be released as a
signal messenger to modify synaptic function, kinase activity, and
protein synthesis.
[0032] Controlled Ca.sup.2+ release from intracellular stores
within a neuron represents an important mechanism for amplifying
Ca.sup.2+ signals received from outside the neuron. Such
intracellular release is also important for the generation of
stimulus-specific spatiotemporal patterning of cytosolic Ca.sup.2+
signals, including Ca.sup.2+ waves, and in switching responses to
low-frequency stimulation from long-term depression to long-term
synaptic potentiation. RyR enables the endoplasmic reticulum to
play an amplifying role in [Ca.sup.2+].sub.i elevation in
neurons.
[0033] Carbonic anhydrase activity, crucial for information coding
and storage, is at least partially activated by intracellular
release of Ca.sup.2+ through the ryanodine receptors (RyR). For
example, the RyR is involved in the GABA-mediated synaptic switch.
The effect of Ca.sup.2+ on carbonic anhydrase appears to be
indirect. In human myelomonocytic cell lines, synthesis of carbonic
anhydrase II is activated by protein kinase C, an effect that is
blocked by 0.1 .mu.m staurosporine. Hormones also regulate the
activity of carbonic anhydrase via cAMP. Thus, the increase in
carbonic anhydrase activity induced by adrenaline and
dibutyryl-cAMP in erythrocytes is enhanced by theophylline, and
phosphorylation by cAMP-dependent protein kinases activates
carbonic anhydrase.
[0034] Carbonic anhydrase plays a crucial role in signal
processing, long term synaptic transformation and attentional
gating of memory storage. There are at least seven isozymes of
carbonic anhydrase in humans. Carbonic anhydrase dysfunction
impairs cognition and is associated with mental retardation,
Alzheimer's Disease and aging. The pharmacological profile of
carbonic anhydrase has been refined and specific activators have
been developed. Carbonic anhydrase, a zinc-containing enzyme,
catalyzes a reversible reaction between CO.sub.2 hydration and
HCO.sub.3.sup.- dehydration. Recent studies indicate that
activation of this enzyme provides a rapid and efficient mechanism
to raise HCO.sub.3.sup.- concentrations in memory-related neural
structures. Increased HCO.sub.3flux through synaptic GABA.sub.A
receptor channels alters postsynaptic neuronal responses to GABA
and thus neuronal responses to diverse signal inputs. In this way,
carbonic anhydrase functions as an effective attentional gate that
controls signal transfer through the neural network. Alterations in
carbonic anhydrase activity in hippocampal CA1 neurons provide a
mechanism for switching between operational states at GABA
releasing synapses, thereby gating signal transfer through the
hippocampal network.
[0035] Because carbonic anhydrase is inactive without zinc at its
active site, it is possible that zinc-containing proteins might
function abnormally in dementia associated with AD and aging. Human
carbonic anhydrase II has a high binding affinity for zinc. Even
concentrations lower than 1 .mu.m are sufficient to induce amyloid
deposits, thus favoring redistribution of zinc from intra to
extracellular sites. The zinc has been shown to induce immediate
aggregation of an N-terminal fragment of .beta.-amyloid
(A.beta..sub.1-40). Zn.sup.21+ is concentrated to .about.1 M in AD
plaques and A.beta. binds to zinc and to deposits at sites that
contain high concentrations of zinc. The hippocampus contains the
highest concentration of zinc in the brain and in AD there is a
decrease in the intracellular concentration of zinc. The essential
role of zinc in initiation of A.beta. formation might explain why
A.beta. deposits are often concentrated in the hippocampus, rather
than distributed uniformly throughout the brain. Reducing A.beta.
formation reduces behavioral impairment in AD transgenic mice
whereas copper-zinc chelators solubilize A.beta. and markedly
reduce A.beta. accumulation in AD transgenic mice. Furthermore,
even if functional zinc deficiency does contribute to AD
pathophysiology, carbonic anhydrase would not be the only protein
affected.
[0036] Carbonic anhydrase inhibitors of the sulfonamide type (e.g.,
topiramate and acetazolamide) are widely used in the treatment of a
variety of disorders such as glaucoma, epilepsy and gastro-duodenal
ulcers. Carbonic anhydrase II deficiency syndrome in humans is also
characterized by renal tubular acidosis, osteoperosis and mental
retardation. Inhibition of carbonic anhydrase reduces and abolishes
acetylcholine-mediated .theta. activity in the hippocampus. Thus,
an important effect of carbonic anhydrase inhibition on hippocampal
function is inhibition of .theta. activity, a synchronized
hippocampal-activity rhythm that is required for spatial memory. In
conscious animals, CNS administration of acetazolamide impairs
spatial learning without affecting other sensory and/or locomotor
behaviors. A single dose of acetazolamide, a specific inhibitor of
carbonic anhydrase, reduces the magnitudes of .theta.-wave
frequency activity measured by electroencephalogram during
rapid-eye-movement sleep by 50% and acute inhibition of carbonic
anhydrase impairs spatial memory.
[0037] Activators of carbonic anhydrase provided an important tool
for the treatment of genetic carbonic anhydrase deficiencies and
memory disorders. Many amines and amino acids (e.g., dopamine,
noradrenaline, adrenaline, histamine, histidine, imidazoles,
phenylalanine or derivatives thereof (See WO 00/56760) and 5-HT)
are carbonic anhydrase activators. Activators of carbonic anhydrase
facilitate the switch between excitatory and inhibitory effects of
GABA.sub.A receptor stimulation that is induced by the temporal
association between activation of cholinergic and GABAergic inputs.
The combination of a methylxanthine (e.g. theophylline) and a
carbonic anhydrase activator show cognitive enhancement of a
specific molecular cascade, which in turn directly affects
attention. The methylxanthine (e.g. theophylline) activates the
ryanodine receptors of the endoplasmic reticulum. Additionally, the
A-1 receptors are antagonized.
[0038] CNS administration of carbonic anhydrase activators (e.g.,
imidazole or phenylalanine) significantly enhances the ability of
rats to learn a water-maze task and to recall the maze from memory.
The present inventors have found that carbonic anhydrase activation
is enhanced when combined with a methylxanthine, with theophylline
being of particularly advantageous. Further discussion of carbonic
anhydrase can be found in Carbonic anhydrase gating of attention:
memory therapy and enhancement; Sun, Miao-Kun and Alkon, Daniel L.,
Trends in Pharmacological Sciences, Vol. 23 No. 2, pp. 83-89
(February 2002), which is hereby incorporated by reference in its
entirety. These spatial-learning effects, which are mediated
through attentional gating of relevant signals in the network, are
sensitive to acetazolamide. Further, training rats in spatial
water-maze task has been found to increase ryanodine receptor
(RyR2) expression in the hippocampus.
[0039] The area of memory and learning impairment is rich in animal
models, which are able to demonstrate different features of memory
and learning processes. (See, for example, Hollister, L. E., 1990,
Pharmacopsychiat., 23, (Suppl II) 33-36). The available animal
models of memory loss and impaired learning involve measuring the
ability of animals to remember a discrete event. These tests
include the Morris Water Maze and the passive avoidance procedure.
In the Morris Water Maze, animals are allowed to swim in a tank
divided into four quadrants, only one of which has a safety
platform beneath the water. The platform is removed and the animals
are tested for how long they search the correct quadrant verse the
incorrect quadrants. In the passive avoidance procedure the animal
remembers the distinctive environment in which a mild electric
shock is delivered and avoids it on a second occasion. A variant of
the passive avoidance procedure makes use of a rodent's preference
for dark enclosed environments over light open ones. Further
discussion can be found in Crawley, J. N., 1981, Pharmacol.
Biochem. Behav., 15, 695-699; Costall, B. et al, 1987,
Neuropharmacol., 26, 195-200; Costall, B. et al, 1989, Pharmacol.
Biochem. Behav., 32, 777-785; Barnes, J. M. et al, 1989, Br. J.
Pharmacol., 98 (Suppl) 693P; Barnes, J. M. et al, 1990, Pharmacol.
Biochem. Behav., 35, 955-962.
[0040] Further data suggest that the inducement of hypoxia and the
damage that would normally result there from can be prevented
through the administration of the combination of a methylxanthine
(e.g. theophylline) and a carbonic anhydrase activator, indicating
that the combination provides a neuroprotective effect. In a
preferred embodiment the carbonic anhydrase activator is
phenylalanine. As a result the present invention could also be used
to treat a variety of conditions including but not limited to AD,
general dementia, mental retardation, "sundown" syndrome, transient
ischemia, and stroke.
[0041] Methylxanthines (i.e., theophylline) are often used to treat
asthmatic conditions. Methylxanthines (i.e., theophylline) are also
known to competitively inhibit phosphodiesterase, the enzyme that
degrades cAMP. An increased concentration of cAMP is proposed to
mediate the observed bronchodilation. Other proposed mechanisms of
action include inhibition of the release of intracellular calcium
and competitive antagonism of the bronchoconstrictor adenosine. The
use of theophylline alone has provided a variety of studies
regarding theophylline's effect on cognition and has generally been
found to be insignificant. (See for example, Weldon, et al.,
Theophylline effects on cognition, behavior, and learning, Arch.
Pediatr. Adolesc. Med., 149(1):90-3 (1995), Newman et al.,
Physiological and neuropsychological effects of theophylline in
chronic obstructive pulmonary disease, Isr. J. Med. Sci.,
30(11):811-6 (1994); Stein et al., Behavioral and cognitive effect
of theophylline: a dose-response study, Ann. Allergy, 70(2):135-40
(1993); Gil et al., Study of the effects of treatment with
theophylline on the cognitive process and behaviour of children
with bronchial asthma, Allergol Immunopathol. 21(5):204-06 (1993);
Fitzpatrick et al., Effect of therapeutic theophylline levels on
the sleep quality and daytime cognitive performance or normal
subjects, Am. Rev. Respir. Dis., 145(6):1355-58 (1992).
[0042] The use of the word, "normal" is meant to include
individuals who have not been diagnosed with or currently display
diminished or otherwise impaired cognitive function. The different
cognitive abilities may be tested and evaluated through known means
well established in the art, including but not limited to tests
from basic motor-spatial skills to more complex memory recall
testing. Non-limiting examples of tests used for cognitive ability
for non-primates include the Morris Water Maze, Radial Maze, T
Maze, Eye Blink Conditioning, Delayed Recall, and Cued Recall while
for primate subjects test may include Eye Blink, Delayed Recall,
Cued Recall, Face Recognition, Minimental, and ADAS-Cog. Many of
these tests are typically used in the mental state assessment for
patients suffering from AD. Similarly, the evaluation for animal
models for similar purposes is well described in the
literature.
[0043] The present compounds can be administered by a variety of
routes and in a variety of dosage forms including those for oral,
rectal, parenteral (such as subcutaneous, intramuscular and
intravenous), epidural, intrathecal, intra-articular, topical and
buccal administration. The dose range for adult human beings will
depend on a number of factors including the age, weight and
condition of the patient and the administration route.
[0044] For oral administration, fine powders or granules containing
diluting, dispersing and/or surface-active agents may be presented
in a draught, in water or a syrup, in capsules or sachets in the
dry state, in a non-aqueous suspension wherein suspending agents
may be included, or in a suspension in water or a syrup. Where
desirable or necessary, flavouring, preserving, suspending,
thickening or emulsifying agents can be included.
[0045] Other compounds which may be included by admixture are, for
example, medically inert ingredients, e.g. solid and liquid
diluent, such as lactose, dextrose, saccharose, cellulose, starch
or calcium phosphate for tablets or capsules, olive oil or ethyl
oleate for soft capsules and water or vegetable oil for suspensions
or emulsions; lubricating agents such as silica, talc, stearic
acid, magnesium or calcium stearate and/or polyethylene glycols;
gelling agents such as colloidal clays; thickening agents such as
gum tragacanth or sodium alginate, binding agents such as starches,
arabic gums, gelatin, methylcellulose, carboxymethylcellulose or
polyvinylpyrrolidone; disintegrating agents such as starch, alginic
acid, alginates or sodium starch glycolate; effervescing mixtures;
dyestuff; sweeteners; wetting agents such as lecithin, polysorbates
or laurylsulphates; and other therapeutically acceptable accessory
ingredients, such as humectants, preservatives, buffers and
antioxidants, which are known additives for such formulations.
[0046] Liquid dispersions for oral administration may be syrups,
emulsions or suspensions. The syrups may contain as carrier, for
example, saccharose or saccharose with glycerol and/or mannitol
and/or sorbitol. In particular syrup for diabetic patients can
contain as carriers only products, for example sorbitol, which do
not metabolise to glucose or which metabolise only a very small
amount to glucose. The suspensions and the emulsions may contain a
carrier, for example a natural gum, agar, sodium alginate, pectin,
methylcellulose, carboxymethylcellulose or polyvinyl alcohol.
[0047] Suspensions or solutions for intramuscular injection may
contain, together with the active compound, a pharmaceutically
acceptable carrier such as sterile water, olive oil, ethyl oleate,
glycols such as propylene glycol and, if desired, a suitable amount
of lidocaine hydrochloride. Solutions for intravenous injection or
infusion may contain a carrier, for example, sterile water which is
generally Water for Injection. Preferably, however, they may take
the form of a sterile, aqueous, isotonic saline solution.
Alternatively, the present compounds may be encapsulated within
liposomes. The present compounds may also utilize other known
active agent delivery systems.
[0048] The present compounds may also be administered in pure form
unassociated with other additives, in which case a capsule, sachet
or tablet is the preferred dosage form.
[0049] Tablets and other forms of presentation provided in discrete
units conveniently contain a daily dose, or an appropriate fraction
thereof, of one of the present compounds. For example, units may
contain from 5 mg to 500 mg, but more usually from 10 mg to 250 mg,
of one of the present compounds.
[0050] In general, one of ordinary skill in the art, acting in
reliance upon personal knowledge and the disclosure of this
application, will be able to ascertain the amounts of these
respective pharmaceutical agents and the amount of the compounds
which should be administered to a subject to achieve the methods
described herein. A "pharmaceutically effective amount," when
referring to a combination of two or more agents, means an amount
of each of the combined agents which is effective in eliciting the
desired biological or medicinal response. For example, the
pharmaceutically effective amount of a composition comprising a
methylxanthine and a carbonic anhydrase inhibitor would be the
amount of a methylxanthine and the amount of a carbonic anhydrase
inhibitor that, when taken together, have a combined effect which
is pharmaceutically effective. In accordance with the methods of
treatment of the present invention, the individual components of
the combination can be administered separately at different times
during the course of therapy or concurrently in divided or single
combination forms. The instant invention is therefore to be
understood as embracing all such regimes of simultaneous or
alternating treatment and the term "administering" is to be
interpreted accordingly.
[0051] It will be appreciated that the pharmacological activity of
the compositions of the invention can be demonstrated using
standard pharmacological models, which are known in the art.
Furthermore, it will be appreciated that the inventive compositions
can be incorporated or encapsulated in a suitable polymer matrix or
membrane for site-specific delivery, or can be functionalized with
specific targeting agents capable of effecting site specific
delivery. These techniques, as well as other drug delivery
techniques are well known in the art.
[0052] All books, articles, or patents references herein are
incorporated by reference to the extent not inconsistent with the
present disclosure. The present invention will now be described by
way of examples, which are meant to illustrate, but not limit, the
scope of the invention.
EXAMPLES
[0053] Water-Maze
[0054] The combination of a methylxanthine and a carbonic anhydrase
activator was tested by administering phenylalanine (50 mg/kg) plus
theophylline to rats 1.5 and 0.5 hours prior to subjecting the rats
to the Morris Water Maze paradigm. Measuring the reduction of the
escape latency in successive trials assessed cognitive ability,
particularly learning. Memory and retention were assessed by
measuring the time spent in the appropriate quadrant a day after
the last trial. The rats treated with the combination of
theophylline and a carbonic anhydrase activator, phenylalanine,
exhibited both faster learning curves and enhanced retention
compared to rats receiving phenylalanine alone or theophylline
alone. (See FIG. 1 graph). Rats treated with the combination of
theophylline and phenylalanine exhibited
[0055] FIG. 2(a) and 2(b) illustrate the swimming time, in each
quadrant of the Morris Water Maze, for control rats and rats
treated with PhePheTheo, respectively. The amount of cumulative
time spent in the target quadrant for treated rats compared to
controls is for the Morris Water Maze shows a significant increase
for treated rats. (See FIG. 2(c)). The results not only demonstrate
that treated rats have improved learning, but that the rats also
demonstrate improved recall.
* * * * *